PROJECT SUMMARY This proposal describes a five-year career development program to prepare Dr. Hongchao Guo for a career as an independent investigator. This program will build on Dr. Guo?s background as a molecular stem cell biologist by providing him expertise in bioinformatics, secretomics, and drug discovery techniques, and knowledge on environmental health, immunology and cardiovascular biology to advance our understanding on how genetic differences in individuals contribute to their susceptibility to smoking-related cardiovascular disease. Dr. Guo will be mentored by Dr. Joseph Wu, Director of Stanford Cardiovascular Institute, who is an expert in cardiovascular disease modeling using induced pluripotent stem cells (iPSCs). In addition, Dr. Guo will be co-mentored by Dr. Kari Nadeau, Director of the Sean N. Parker Center for Allergy and Asthma Research at Stanford University, who is an expert in studying aerosolized pollutants including tobacco smoke on the development of immune dysfunction in primary immune disease, allergy and asthma. The K99 phase of Dr. Guo?s training will consist of structured mentorship by the primary mentor and co-mentor, close interactions with collaborators, complementary meeting with advisory committee members, a provocative research project, and a tailored program for career development and transition. In Dr. Guo?s previous works, he has recapitulated key features of nicotine-induced vascular dysfunction in patients using patient-specific iPSC-derived endothelial cell (iPSC- EC). He has also shown that genetic variants in nicotinic acetylcholine receptor exacerbated nicotine-induced EC dysfunction via increasing inflammation cytokines expression and apoptosis, which allow for analysis of the molecular mechanism in iPSC-EC model with a level of depth and resolution never before achieved. With the current advancement in high-throughput RNA sequencing and the cutting-edge secretomics and drug screening technologies, Dr. Guo is in a unique position to stratify the patient risk of genetic variants in nicotinic receptors for smoking-induced vascular diseases, and to study the potential mechanisms, with the ultimate goal to discover biomarkers and precise treatment for cardiovascular diseases and risks. In the K99 phase, Dr. Guo will generate and characterize patient-specific and isogenic iPSC-EC models for studying the susceptibility of three different nAChR variants to nicotine-induced vascular dysfunction (Aim 1). With the platform, Dr. Guo will then integrate high-throughput RNA sequencing and cutting-edge secretomics technologies to define the key molecular basis and secretomic biomarkers for the risk of these variants to smoking-mediated vascular diseases (Aim 2). In the R00 phase, Dr. Guo will develop a screening platform to examine the beneficial effect of anti-inflammatory drugs and screen mechanism-oriented small molecules in iPSC-EC carrying these risk variants (Aim 3). Collectively, Dr. Guo?s proposed work will create a valuable platform to evaluate the risk of these variants to smoking-related vascular disease and to reveal the molecular mechanisms and biomarkers. Additionally, this work will lead to studying of mechanism-oriented treatment, which will be carried out by Dr. Guo as an independent investigator.